Cancer Death In American Women Research Articles

Abstract 4492: Deferiprone optimization for the treatment of triple-negative breast cancer

Abstract Triple-negative breast cancer (TNBC) is the leading cause of new cancer cases and the second leading cause of cancer deaths in American women disproportionately affecting Black and Hispanic women under 40. Currently, there is no targeted therapy for TNBC and treatment options to manage this lethal disease include surgery, adjuvant chemotherapy and radiotherapy. Among the chemotherapy agents, combination of the PARP inhibitors with DNA-damaging results in a more promising anti-tumor effect. A better understanding of TNBC etiology has aided in unraveling the roles of two additional cellular targets - androgen receptor (AR) and histone lysine demethylase (KDM) - in the viability and survival of TNBC. Interestingly, AR overexpression is the most important driver of prostate cancer (PCa). A class of current medications used to treat PCa act as androgen receptor (AR) antagonists (or antiandrogens). Antiandrogens are under investigation in the clinic as potential therapies for luminal AR (LAR) subtypes of TNBC, a particularly worrisome subtype that frequently metastasizes to regional lymph nodes and bones. Although LAR TNBC constitutes 15-20% of TNBCs, it is characterized by minimal response to chemotherapy and aggressiveness. It is therefore plausible to repurpose and optimize antiandrogens, in the context of designed multiple ligands; to obtain a TNBC-targeting therapy effective against several subtypes of this heterogenous disease, including the basal subtype that is responsible for ~80% of TNBC incidence. The proposed therapeutics are dual-acting antiandrogen-KDM inhibitors. The antiandrogen moiety of these agents will enable TNBC cell-targeting and translocation of the drug into the nucleus where inhibition of KDM will prevent transcription of KDMs that promote tumor growth. The three pharmacophores of these agents are (i) an AR binding group, (ii) a linker, and (iii) an iron-binding group. Two functional classes of these compounds, alkyl- and aryl-derivatives, vary in the design of the AR binding moiety. To optimize these agents, the methylene linker length is varied in each class for in vitro determination of the optimal length for increased drug efficacy. The compounds are tested on TNBC cell lines (MDA-MB-231, low AR+; and MDA-MB-453, high AR+), and ER+ BCa (MCF-7) while PCa cell lines (LNCaP PCa, AR+; and DU 145, PCa, AR-) serve as controls for AR dependency of the compounds. Longer linker length compounds exhibited low micromolar cytotoxicity and selectivity for AR+ TNBC and PCa cell lines. Further analysis of lead compounds in TNBC cells aided in understanding the effects on targets, transcriptomic effects via RNA sequencing analysis and the mechanism of action. Thus far, the designed dual-acting therapeutics have potential to revolutionize the design of anticancer drugs and improve treatment outcome for TNBC patients. Citation Format: Alexis Johnston, Adegboyega K. Oyelere. Deferiprone optimization for the treatment of triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4492.

Epigenetic modulation of antitumor immunity and immunotherapy response in breast cancer: biological mechanisms and clinical implications.

Breast cancer (BC) is the most common non-skin cancer and the second leading cause of cancer death in American women. The initiation and progression of BC can proceed through the accumulation of genetic and epigenetic changes that allow transformed cells to escape the normal cell cycle checkpoint control. Unlike nucleotide mutations, epigenetic changes such as DNA methylation, histone posttranslational modifications (PTMs), nucleosome remodeling and non-coding RNAs are generally reversible and therefore potentially responsive to pharmacological intervention. Epigenetic dysregulations are critical mechanisms for impaired antitumor immunity, evasion of immune surveillance, and resistance to immunotherapy. Compared to highly immunogenic tumor types, such as melanoma or lung cancer, breast cancer has been viewed as an immunologically quiescent tumor which displays a relatively low population of tumor-infiltrating lymphocytes (TIL), low tumor mutational burden (TMB) and modest response rates to immune checkpoint inhibitors (ICI). Emerging evidence suggests that agents targeting aberrant epigenetic modifiers may augment host antitumor immunity in BC via several interrelated mechanisms such as enhancing tumor antigen presentation, activation of cytotoxic T cells, inhibition of immunosuppressive cells, boosting response to ICI, and induction of immunogenic cell death (ICD). These discoveries have established a highly promising basis for using combinatorial approaches of epigenetic drugs with immunotherapy as an innovative paradigm to improve outcomes of BC patients. In this review, we summarize the current understanding of how epigenetic processes regulate immune cell function and antitumor immunogenicity in the context of the breast tumor microenvironment. Moreover, we discuss the therapeutic potential and latest clinical trials of the combination of immune checkpoint blockers with epigenetic agents in breast cancer.

Abstract 3092: Optimization of deferiprone for the treatment of triple-negative breast cancer

Abstract Triple-negative breast cancer (TNBC) is the leading cause of new cancer cases and the second leading cause of cancer deaths in American women. TNBC is one of the most difficult cancers to treat due to the lack of three receptors, estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor (HER2), found in regular forms of breast cancer (BCa). Currently, there is no targeted therapy for TNBC and treatment options to manage this lethal disease include surgery, adjuvant chemotherapy and radiotherapy. A better understanding of TNBC etiology has aided in unraveling the roles of two additional cellular targets - androgen receptor (AR) and histone lysine demethylase (KDM) - in the viability and survival of TNBC. Interestingly, AR overexpression is the most important driver of prostate cancer (PCa). A class of current medications used to treat PCa act as androgen receptor (AR) antagonists (or antiandrogens). It is therefore plausible to repurpose and optimize these antiandrogens for TNBC therapy. The proposed therapeutics are dual-acting antiandrogen-KDM inhibitors. The antiandrogen moiety of these agents will enable TNBC cell-targeting and translocation of the drug into the nucleus where inhibition of KDM will prevent transcription of KDMs that promote tumor. The three pharmacophores of these agents are (i) a surface recognition cap, (ii) a linker, and (iii) an Fe2+-binding group. Two classes of these compounds, alkyl-derivative and benzyl-derivative, vary in the design of the surface recognition cap. To optimize these agents, the linker length is varied from 2-7 methylene groups in each class and the optimal length will be determined through in vitro studies. The compounds are tested on five cancer cell lines: TNBC cell lines (MDA-MB-231, low AR+; and MDA-MB-453, high AR+), and ER+ BCa (MCF-7) while PCa cell lines (LNCaP PCa, AR+; and DU 145, PCa, AR-) serve as controls for AR dependency of the effects of test compounds. Longer linker length compounds exhibited low micromolar cytotoxicity and selectivity for MDA-MB-231 (TNBC) and LNCaP (PCa) cell lines due to the presence of the AR. Western blot analysis on lead compounds supported significant downregulation of AR in MDA-MB-231 cells confirming the predicted mechanism of action. Thus far, the designed dual-acting therapeutics have potential to revolutionize the design of anticancer drugs and improve treatment outcome for TNBC patients. Citation Format: Alexis Johnston, Adegboyega K. Oyelere. Optimization of deferiprone for the treatment of triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3092.

Abstract P1-05-34: MicroRNAs as Prognostic Markers in Breast Cancer

Abstract Background: Breast cancer (BC) is the most common cancer and the second leading cause of cancer death in American women. BC disproportionally affects women of ethnic minorities, especially Black and Hispanic. While this disparity has been acknowledged and social determinations of health remains an important contributor, BC pathophysiology is yet to be elucidated in minorities. MicroRNAs (miRNA), including but not limited to let-7, 125-3p, 192-5p, 451a, play regulatory roles in cancer pathobiology and progression in many cancer types, including but not limited to BC. In our lab, in vitro and in vivo studies on mouse models demonstrated that miR-23b and miR-27b had crucial roles in cancer progression in BC via mechanisms that involves nischarin, an integrin-binding protein, and were associated with worse outcome. Moreover, miRNAs can be detected and are stable in blood samples, and thus the detection has a profound clinical impact on diagnosis. Study Design: Between April 2021 and May 2022, 30 newly diagnosed patients with BC and 30 patients without any cancer diagnosis at University Medical Center- New Orleans were enrolled on the study (IRB# 936). 86% of patients are African American or Hispanics. Plasma samples were collected before any medical intervention for BC. Plasma miRNAs were isolated and miRNA expression were performed utilizing RT-PCR. Results: Expressions of let-7 and 125a miRNAs are two times higher in plasma of patients with BC compared to patients without a diagnosis of cancer. There was no difference in miRNA expression in circulatory 23b, 27b, 192-5p, and 451a. Among breast cancer types, there are two-fold decreased expression of 192-5p and 451a in triple negative compared to hormone positive BC. Conclusion: This project will add to the body of knowledge on roles of miRNAs as prognostic markers in patients with BC, more importantly, triple negative BC, and elucidate pathophysiologic mechanism contributing to racial disparity in breast cancer treatment and recurrence. In the near future, we aim to examine effects of chemotherapy and surgery on the expression of circulatory microRNAs. Citation Format: Anh Q. Nguyen, Colin Cunningham, Samuel Okpechi, Hassan Yousefi, Shawn McKinney, Brian Boulmay, Agustin Garcia, Suresh Alahari. MicroRNAs as Prognostic Markers in Breast Cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P1-05-34.

Molecular aberrations in late-stage dedifferentiated/undifferentiated endometrial carcinoma: A review of current literature.

e17632 Background: Uterine cancer remains the fourth most common cancer diagnosed and seventh most common cause of cancer death in American women. Four molecular subtypes have been created to categorize and prognosticate endometrial carcinoma based on their molecular aberrations, but are often based on initial tumor presentation and are associated with high cost to the healthcare system. Methods: A comprehensive literature search was performed on PubMed, ClinicalTrials.gov, Embase, and WebofScience. Abstracts and publications from ASCO/ASH were also included. Results: 32 articles were included in our systematic review. A total of 388 women were analyzed in the main group analysis and nearly half of the patients were diagnosed as stage III or IV (n = 178, 46%). MSI was present in 66% of all tumors tested and PD-L1 was overexpressed in 78%. Eighty-four percent (84%) of the tumors were negative for ER, up to 44% had loss in SWI/SNF complex, and 64% lost expression of e-cadherin. Pax-8 and fascin proteins were over expressed in 29% and 88% of patients tested, respectively. In a sub-group analysis, evidence of progression and use of adjuvant radiotherapy were significantly associated with a later stage of diagnosis ( p = 0.052, 0.044). Conclusions: We found no statistically significant association between stage of diagnosis and unique mutation. Patients with evidence of progression or recurrence were found to have disease in atypical locations. Unique mutations which categorize patients at time of diagnosis may be different upon progression or recurrence of the tumor. While a novel classification system has been produced to assist with prognosis and treatment guidelines, these tests are not widely available in the community hospital setting.[Table: see text]

Abstract 6403: Characterization of small molecule inhibitors of PELP1 for treating advanced breast cancer

Abstract Background: Breast cancer (BC) is the most commonly diagnosed cancer and the second leading cause of cancer death in American women. BC is composed of distinct molecular subtypes, such as ER positive BC (ER+) and triple negative BC (TNBC). Development of novel effective therapies for patients with therapy resistant BC (TR-BC) and TNBC remains the highest unmet need in patient treatment and survivorship. Proline, glutamic acid-, and leucine-rich protein 1 (PELP1) plays a critical role in multiple nuclear receptor functions leading to TR-BC and TNBC progression. PELP1 expression is dysregulated in BC, is a prognostic indicator of poorer BC survival, and its deregulation contributes to TR-BC. The objective of this study is characterization of a small molecule inhibitor of PELP1 (SMIP34) as novel therapeutic for treating advanced BC. Methods: Yeast two-hybrid screening was used to identify PELP1 Inhibitory Peptide 1 (PIP1). Direct binding of PIP1 to PELP1 was confirmed using biotin pull-down assays and inhibition of BC proliferation confirmed using MTT assays. Hit-Ligand interaction site with PIP1 hot spot residues based on 3D alignment and morphology was used to generate a library of small chemical molecules that function as peptidomimetics of PIP1. In vitro activity was assessed using CellTiter Glo, MTT, colony formation and matrigel invasion assays in multiple BC models. Mechanistic studies were conducted using Western blot, and reporter gene assays. Xenograft and patient derived explant (PDEX) assays were used for preclinical evaluation. Results: Screening utilizing MTT assays lead to the selection of SMIP34 as an inhibitor of PELP1. SMIP34 treatment reduced cell viability at an IC50 of 3-10µM in a panel of BC cells. Additionally, SMIP34 showed no activity in human mammary epithelial cells. Knockdown of PELP1 in BC cells significantly reduced the SMIP34 activity, confirming target specificity. Furthermore, SMIP34 treatment significantly reduced the invasiveness and colony formation of TR-BC and TNBC cells. Mechanistic studies using Western blot analysis confirmed that SMIP34 binding to PELP1 contributes to its degradation. In combination studies, SMIP34 displayed synergy and enhanced the efficacy of current chemotherapeutics Cisplatin and Paclitaxel. In PDEX assays, SMIP34 significantly decreased tumor proliferation as measured by Ki67 staining. In xenograft models, SMIP34 (10mg/kg/s.c.) treatment resulted in significant reduction in tumor volume compared to vehicle control. Furthermore, overall mouse body weight in both control and SMIP34 treated groups were similar, suggesting no overt signs of toxicity. Conclusion: We have developed a first-in-class small molecule inhibitor of PELP1 (SMIP34) displaying therapeutic efficacy against TR-BC and TNBC in vitro, ex vivo, and in vivo. Supported by CPRIT Predoctoral Fellowship CPRIT RTA; RP170345 (K.A. Altwegg) and VA grant I01BX004545 (R.K.V) Citation Format: Kristin Ann Altwegg, Suryavathi Viswanadhapalli, Monica Mann, Uday P. Pratap, Mengxing Li, Junhao Liu, Yiliao Luo, Gangadhara R. Sareddy, Hariprasad Vankayalapati, Ratna K. Vadlamudi. Characterization of small molecule inhibitors of PELP1 for treating advanced breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6403.

Abstract 4246: Characterizing breast cancer health disparities through molecular and genomic profiling of a diverse breast cancer cohort

Abstract The major indicators of breast cancer health disparities may include higher incidence, earlier onset, faster progression, and poorer outcomes. The ultimate goal of health disparities research is to elucidate actionable mechanisms of these differences to define the means of intervention that will help reduce the disparity. Breast cancer is the most common cancer and second leading cause of cancer death in American women and the new cases of invasive cancer have been increased lately in the United State. It is a heterogeneous disease with few modifiable risk factors, characterized by multiple subtypes that influence different populations with varying incidence, morbidity and mortality. Though recent genomic advances have enabled the stratification of breast cancer into numerous subtypes with differing therapeutic and prognostic values, it remains unclear why the incidence and outcome of certain breast cancers show an uneven distribution across different race/ethnic populations. For example, the higher frequency of more aggressive forms of breast cancer in young women of African descent is well known, yet why this occurs remains to be determined. To understand the mechanisms of breast cancer health disparity, we performed a molecular and genomic profiling on a unique cohort of racially diverse breast cancer patients from a catchment area in the northern part of east North Carolina and found earlier onset of breast cancer in women of African Ancestry (AA) with 32.1 % occurring before age 50 compared to 22.5 % for women of European Ancestry (EA). Furthermore, women of AA have higher levels of obesity with 35.5 % having a Body mass index (BMI) above 35 compared to 17.9 % for women of EA. Notably, women of AA also showed presence of higher stage of breast cancer with more than 26 % having stage 3 or 4 compared to 16.5 % for women of EA. We also found that women of AA showed poorer overall survival outcome from breast cancer compare to EA with Hazard Ratio of 1.67. The panel of exome sequencing of 147 tumors from this cohort revealed differential mutational frequencies in African Ancestry versus European Ancestry. The insertion/deletion mutated genes found only in African Ancestry included PAX5, CYP2A7, UGT2B17, CYP4A11, ACAA1, KIT and NAT2 with high percentage of cases with mutation account, whereas those genes unique to European Ancestry included GSTP1, PRSS53, AURKA, ATF1, BIRC2, PGAP3 and GSTA4. In addition to this, there are differentially mutated genes across specific breast cancer subtypes where ABL2, UBA1, CYP4F8, PIK3CG, CTNNA1, CYP39A1 genes are only associated with Triple Negative Breast Cancer patients. By characterizing the risk of breast cancer among ethnically diverse groups, we provide needed insights that will not only improve the development of personalized prognostic and predictive biomarkers but may also enhance breast cancer screening, treatment, survival and prevention. Citation Format: Jung S. Byun, Tingfen Yan, Adam Sowalsky, Jack Zhu, Paul Meltzer, Nasreen Vohra, Peter Kragel, Anna Napoles, Eliseo Perez-Stable, Kevin Gardner. Characterizing breast cancer health disparities through molecular and genomic profiling of a diverse breast cancer cohort [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4246.

Abstract 604: Metabolomics in predicting breast cancer treatment responses

Abstract Breast cancer is the most frequently diagnosed cancer in women and the second leading cause of cancer death in American women. Postoperative adjuvant radiotherapy (RT) significantly improve local-regional recurrence and survival. Therefore, there has been increasing usage of adjuvant RT in early-stage breast cancer patients. Although well tolerated by some patients, 48% patients in our study experience RT-induced early adverse skin reactions (EASRs) that negatively impact quality of life. Therefore, we evaluated urine metabolomics in predicting RT-induced EASRs and precision intervention. Based on the pilot data of 478 metabolites in 60 patients, we used the MetaboAnalyst 3.0 (www.metaboanalyst.ca) to conduct metabolomic data analysis, visualization and interpretation. Using KEGG pathway analysis, we have identified 7 pathways that are significantly associated with RT-induced EASRs (i.e., False Discovery Rate (FDR) p less than 0.05). The alanine, aspartate and glutamate metabolism pathway has the most significant FDR p-value and the highest impact value of 0.60 in predicting RT-induced EASRs. In summary, to the best of our knowledge, this is the first study in breast cancer patients associating glutamate metabolism pathway to RT-induced EASRs. Our research will have significant clinical impact because: (i) many tumor cells depend on glutamine-glutamate as energy source for growth and survival, (ii) breast cancer cells secrete high levels of glutamate often metastasize to bone with severe pain issue, (iii) glutaminase inhibitors enhance chemotherapy in triple negative breast cancer, (iv) two thirds of cancer patients receive RT, and (v) glutamate metabolism is associated with RT-induced EASRs. With increasing recent interest in targeting tumor metabolism, we anticipate that glutaminase inhibitors will also have application in preventing RT-induced EASRs in addition to their anticancer activities in triggering metabolic crisis in tumor cells and enhancing chemotherapy. Citation Format: Carolina Puyana Barcha, Eunkyung Lee, Cristiane Takita, Jean L. Wright, Jennifer Hu. Metabolomics in predicting breast cancer treatment responses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 604.

Practice Bulletin No. 179 Summary: Breast Cancer Risk Assessment and Screening in Average-Risk Women.

Breast cancer is the most commonly diagnosed cancer in women in the United States and the second leading cause of cancer death in American women (1). Regular screening mammography starting at age 40 years reduces breast cancer mortality in average-risk women (2). Screening, however, also exposes women to harm through false-positive test results and overdiagnosis of biologically indolent lesions. Differences in balancing benefits and harms have led to differences among major guidelines about what age to start, what age to stop, and how frequently to recommend mammography screening in average-risk women (2-4).Breast cancer risk assessment is very important for identifying women who may benefit from more intensive breast cancer surveillance; however, there is no standardized approach to office-based breast cancer risk assessment in the United States. This can lead to missed opportunities to identify women at high risk of breast cancer and may result in applying average-risk screening recommendations to high-risk women. Risk assessment and identification of women at high risk allow for referral to health care providers with expertise in cancer genetics counseling and testing for breast cancer-related germline mutations (eg, BRCA), patient counseling about risk-reduction options, and cascade testing to identify family members who also may be at increased risk.The purpose of this Practice Bulletin is to discuss breast cancer risk assessment, review breast cancer screening guidelines in average-risk women, and outline some of the controversies surrounding breast cancer screening. It will present recommendations for using a framework of shared decision making to assist women in balancing their personal values regarding benefits and harms of screening at various ages and intervals to make personal screening choices from within a range of reasonable options. Recommendations for women at elevated risk and discussion of new technologies, such as tomosynthesis, are beyond the scope of this document and are addressed in other publications of the American College of Obstetricians and Gynecologists (ACOG) (5-7).

Practice Bulletin Number 179: Breast Cancer Risk Assessment and Screening in Average-Risk Women.

Breast cancer is the most commonly diagnosed cancer in women in the United States and the second leading cause of cancer death in American women (1). Regular screening mammography starting at age 40 years reduces breast cancer mortality in average-risk women (2). Screening, however, also exposes women to harm through false-positive test results and overdiagnosis of biologically indolent lesions. Differences in balancing benefits and harms have led to differences among major guidelines about what age to start, what age to stop, and how frequently to recommend mammography screening in average-risk women (2-4).Breast cancer risk assessment is very important for identifying women who may benefit from more intensive breast cancer surveillance; however, there is no standardized approach to office-based breast cancer risk assessment in the United States. This can lead to missed opportunities to identify women at high risk of breast cancer and may result in applying average-risk screening recommendations to high-risk women. Risk assessment and identification of women at high risk allow for referral to health care providers with expertise in cancer genetics counseling and testing for breast cancer-related germline mutations (eg, BRCA), patient counseling about risk-reduction options, and cascade testing to identify family members who also may be at increased risk.The purpose of this Practice Bulletin is to discuss breast cancer risk assessment, review breast cancer screening guidelines in average-risk women, and outline some of the controversies surrounding breast cancer screening. It will present recommendations for using a framework of shared decision making to assist women in balancing their personal values regarding benefits and harms of screening at various ages and intervals to make personal screening choices from within a range of reasonable options. Recommendations for women at elevated risk and discussion of new technologies, such as tomosynthesis, are beyond the scope of this document and are addressed in other publications of the American College of Obstetricians and Gynecologists (ACOG) (5-7).

Abstract 3279: Circulating inflammatory biomarker C-reactive protein and radiotherapy-related pain in breast cancer patients

Abstract Background: Breast cancer is the most frequently diagnosed cancer and the second leading cause of cancer death in American women. Post-surgery adjuvant radiotherapy (RT) significantly improves clinical outcomes. However, many patients develop treatment-related pain that negatively impacts quality of life. Methods: We evaluated an inflammatory biomarker, C-reactive protein (CRP) in RT-associated pain in a prospective study of 366 breast cancer patients (64% Hispanic whites, 20% African Americans, 16% non-Hispanic whites) undergoing RT. In each patient, we measured pre- and post-RT plasma CRP levels using a highly-sensitive ELISA kit. Pain score was assessed at pre- and post-RT as the mean of four pain severity items (i.e., pain at its worst, least, average, and now) from the Brief Pain Inventory. Pain scores of 4-10 were considered clinically-relevant pain. Association between pain and CRP levels was evaluated using logistic regression models after adjusting for potential confounders. Results: Overall, 16% patients at pre-RT and 31% patients at post-RT had pain. In multivariable analyses, RT-related pain score change >4 was significantly associated with CRP change >1 mg/L (OR=3.24, 95%CI=1.33, 7.88), age <50 (OR=3.27, 95%CI=1.20, 8.92), and 3+ comorbidity conditions (OR=5.91, 95%CI=1.33, 26.82). Conclusion: This is the first study to suggest that increased inflammatory biomarker, CRP level was associated with RT-related pain. Impact: Future larger studies are warranted to validate our findings and facilitate the discovery and development of anti-inflammatory agents to protect patients from RT-related pain and improve quality of life in breast cancer patients undergoing RT. Citation Format: Eunkyung Lee, Omar L. Nelson, Carolina Puyana, Cristiane Takita, Jean L. Wright, Wei Zhao, Isildinha M. Reis, Rick Lin, Jennifer J. Hu. Circulating inflammatory biomarker C-reactive protein and radiotherapy-related pain in breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3279. doi:10.1158/1538-7445.AM2017-3279

Abstract 1053: Synergistic anticancer effects of combined γ-tocotrienol and pterostilbene is associated with a suppression in Rac1/WAVE 2 signaling in highly malignant breast cancer cells

Abstract Breast cancer is the most common cancer and the second leading cause of cancer death in American women. Rho GTPases play a crucial role in regulating different cellular functions, including cell proliferation, gene expression, cell morphology, actin polymerization, cell motility, metastasis and apoptosis. Wiskott-Aldrich syndrome protein family verprolin-homologous protein2 (WAVE2) is a member within the Wiskott-Aldrich syndrome protein (WASP) family displays a significant role in actin polymerization and cytoskeletal organization which, are essential for cellular migration and invasion. γ-Tocotrienol is a natural isoform within the vitamin E family that displays anticancer activity against a variety of cancer cell types. Pterostilbene is a natural dimethyl ether analog of resveratrol that has several beneficial effects such as anticancer activity. Studies were conducted to examine the effects of combined γ-tocotrienol and pterostilbene on Rac1/WAVE 2 signaling in mouse +SA and human MDA-MB231 breast cancer cells and cellular migration. Results show that combined treatment of γ-tocotrienol and pterostilbene caused a synergistic inhibition of both mouse +SA and human MDA-MB231 breast cancer cells growth, and corresponding reduction in Rac1/WAVE2 signaling as characterized by a significant inhibition in the levels of phospho-Rac1/cdc42, WAVE2, Arp2, and Arp3 expression. Additional studies indicated that this combination resulted in a significant inhibition in mammary cancer cell migration. In summary, these findings strongly suggest that combined treatment of γ-tocotrienol with pterostilbene may provide great benefit as a therapeutic approach in the treatment of metastatic breast cancer. Supported by a grant from the Louisiana Cancer Foundation. Citation Format: Ibrahim G. Algayadh, Paul William Sylvester. Synergistic anticancer effects of combined γ-tocotrienol and pterostilbene is associated with a suppression in Rac1/WAVE 2 signaling in highly malignant breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1053. doi:10.1158/1538-7445.AM2017-1053

Berberine Reverses Hypoxia-induced Chemoresistance in Breast Cancer through the Inhibition of AMPK- HIF-1α.

Breast cancer is the most common type of cancer and the second leading cause of cancer death in American women. Chemoresistance is common and inevitable after a variable period of time. Therefore, chemosensitization is a necessary strategy on drug-resistant breast cancer. In this study, MCF-7 breast cancer cell was cultured under hypoxia for a week to induce the resistance to doxorubincin (DOX). The effect of different doses of berberine, a traditional Chinese medicine, on DOX sensitivity to MFC-7/hypoxia cells was observed. We found that hypoxia increased DOX resistance on breast cancer cells with the AMPK activation. Low-dose berberine could resensitize DOX chemosensitivity in MCF-7/hypoxia cell, however, high-dose berberine directly induced apoptosis. The intriguing fact was that the protein expressions of AMPK and HIF-1α were down-regulated by berberine, either low dose or high dose. But the downstream of HIF-1α occurred the bifurcation dependent on the dosage of berberine: AMPK-HIF-1α-P-gp inactivation played a crucial role on the DOX chemosensitivity of low-dose berberine, while AMPK-HIF-1α downregulaton inducing p53 activation led to apoptosis in high-dose berberine. These results were consistent to the transplanted mice model bearing MCF-7 drug-resistance tumor treated by berberine combined with DOX or high-dose berberine alone. This work shed light on a potentially therapeutic attempt to overcome drug-resistant breast cancer.

Abstract 3357: Breast cancer-associated a2 isoform vacuolar ATPase regulates neutrophils: potential association for tumor invasion

Abstract Breast cancer is the most common cancer worldwide and it is a prominent cause of cancer death in developing countries and the second leading cause of cancer death in American women. Breast cancer infiltrating immune cells like macrophages and neutrophils, TAM and TAN respectively are highly influencing tumor progression. Factors modulating TAN to produce pro-tumorigenic factors are not well understood. Vacuolar ATPases are overexpressed on many tumors and their expression highly correlates to the tumors metastatic potential. In tumor cells a cleaved peptide from the N-terminal domain of the a2-Isoform vacuolar ATPase (a2NTD) is secreted in the microvesicles and promotes pro-tumorigenic properties of macrophages as in TAM. In the current study we show by immunofluorescent staining that human breast cancer cell lines MDA-MB-231 and MCF-7 overexpress a2V in comparison to the non-tumorigenic cell line MCF-10A. Also, immunohistochemical analysis showed that a2NTD is highly expressed on infiltrative ductal carcinoma (IDC, represents 80% of all breast cancers) sections as compared to normal breast tissue. In order to investigate the regulatory effect of a2NTD on neutrophils we stimulated freshly isolated neutrophils from healthy volunteers with recombinant a2NTD. a2NTD treatment up-regulates the gene expression of anti-inflammatory mediators as IL-10 and interleukin 1 receptor antagonist (IL-1RA) and pro-inflammatory cytokines and chemokines like TNFα, IL-1β, IL-1α, IL-6, CCL-2, CXCL-1 and CXCL-2 in neutrophils. The multiplex luminex assay was done confirming the up-regulation of these mediators at the protein level. Moreover, a2NTD stimulated neutrophils enhanced the secretion of angiogenic and metastatic factors such as IL-8, VEGF, MMP-9 and elevated the gelatinase activity of MMP-9, which was detected by zymography. To evaluate the influence of a2NTD treated neutrophils on tumor progression we did an in vitro angiogenesis assay using Human Umbilical Vein Endothelial Cells (HUVECs). a2NTD treated neutrophils supernatant caused a 4 fold increase in the number of the capillary tubes formed by HUVECs and increased the branching of these tubes as well as the formation of closed structures. Furthermore, we tested the potential of a2NTD treated neutrophils on the invasiveness of the MDA-MB231 and MCF-7 breast cancer cell lines using in vitro trans-well invasion assay. Interestingly, a2NTD treated neutrophils supernatant promoted the MDA-MB-231 and MCF-7 invasion by 20 and 8 percent respectively as compared with untreated neutrophil supernatant. These data demonstrate the potential effect of tumor associated a2V and its soluble peptide a2NTD in regulating the sterile inflammation present in tumor microenvironment by modulating neutrophils, which promotes angiogenesis and tumor invasion. Citation Format: Safaa A. Ibrahim, Magdy Amin, Arpita Kulshrestha, Sahithi Pamarthy, Kenneth D. Beaman. Breast cancer-associated a2 isoform vacuolar ATPase regulates neutrophils: potential association for tumor invasion. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3357. doi:10.1158/1538-7445.AM2015-3357

Abstract 3327: Metabolomics: Potential molecular mechanisms of radiotherapy (RT)-related normal tissue toxicities in breast cancer

Abstract Breast cancer is the most frequently diagnosed cancer in women and the second leading cause of cancer death in American women. Clinical guidelines recommend that when breast-conserving surgery is provided as a primary therapy for early-stage breast cancer, RT should follow to improve clinical outcome. Although well tolerated by most patients, even with improved RT technology, breast cancer patients experience moist desquamation as early adverse skin reactions. To elucidate the molecular mechanisms involved in RT-related normal tissure toxicity, global metabolomic profiles were determined in urine samples comparing across paired pre-RT and post-RT samples from 60 breast cancer patients exhibiting low (n = 30) or high (n = 30) skin toxicity in response to RT. The extracted samples were split into equal parts for analysis on the GC/MS and LC/MS platforms. The dataset comprises a total of 478 compounds of known identity. Preliminary data suggest that pre-RT glucose and a variety of derivatives linked to the pentose phosphate pathway, such as ribulose, or the consumption of pyruvate in the mitochondrial tricarboxylic acid cycle, including citrate and succinate, were significantly elevated in the high toxicity group. Multiple metabolites derived from the oxidation of aromatic compounds such as amino acids and dietary polyphenolics - vanillate, 3-hydromandelate, tyramine, 3-methoxy-4-hydroxyphenylglycol, and 2-hydroxyphenylacetate - were elevated in a trending or significant fashion at the pre-RT baseline in the high toxicity relative to the low toxicity group. Pre-RT ascorbate depletion and accumulation of tobacco metabolites were associated with radiation sensitivity. Ascorbate was significantly decreased at baseline in the high toxicity group and its pre-RT levels were negatively correlated with post-RT skin toxicity. Nicotine derivatives were only detected in 12% of the pre- and post-RT samples but their presence seemed to be a risk factor that could predispose smokers to unfavorable outcomes during radiation therapy. Histamine breakdown markers provided an indication of a heightened inflammatory activity in individuals who experienced toxicity as suggested that anti-histamines might be useful to prevent sensitivity. Our preliminary data suggest the potential application of metabolomics in predicting and monitoring RT-related normal tissue toxicities; future larger studies are warranted to confirm out promising findings and identify targeted metabolic pathways for effective intervention. Citation Format: Jennifer J. Hu, Eunkyung Lee, Cristiane Takita, Jean L. Wright, Omar L. Nelson. Metabolomics: Potential molecular mechanisms of radiotherapy (RT)-related normal tissue toxicities in breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3327. doi:10.1158/1538-7445.AM2015-3327

Colorectal cancer screening and prevention in women.

Colorectal cancer (CRC) is one of the leading cancers and cause of cancer deaths in American women and men. Females and males share a similar lifetime cumulative risk of CRC however, substantial differences in risk factors, tumor biology, and effectiveness of cancer prevention services have been observed between them. This review distills the evidence documenting the unique variation observed between the genders relating to CRC risk factors, screening and prevention. Consistent evidence throughout the world demonstrates that women reach equivalent levels of adenomas and CRC as men but it occurs nearly a decade later in life than in their male counterparts. Women have a higher proportion of tumors which are hypermethylated, have microsatellite instability and located in the proximal colon suggesting the serrated pathway may be of greater consequence in them than in men. Other CRC risk factors such as smoking, diet and obesity have been shown to have disparate effects on women which may related to interactions between estrogen exposure, body fat distribution, and the biologic underpinnings of their tumors. There is data showing the uptake, choice, and efficacy of different CRC screening methods in women is dissimilar to that in men. The mortality benefit from FOBT, sigmoidoscopy, and protection from interval CRC by colonoscopy appears to be lower in women than men. A greater understanding of these gender idiosyncrasies will facilitate an personalized approach to CRC prevention and should ultimately lead to a reduced burden of disease.

Making women whole again: a review of breast reconstruction.

Breast cancer is the second most common type of cancer and the second leading cause of cancer death in American women. Fewer than 25% of women diagnosed with breast cancer are aware of all of their reconstructive options. This article reviews options for breast reconstruction and the team approach that can help women feel whole again after breast cancer surgery.

Abstract 2010: SPHK1 promotes lung metastasis of breast cancer

Abstract Breast cancer is the second most commonly diagnosed cancer and second leading cause of cancer death in American women. The majority of cancer-related fatalities are a result of metastases to distant organs, rather than due to the primary tumors themselves. The role of the lipid metabolic pathways in breast cancer progression and metastasis has become a focal point of interest in recent years and our work is mainly focused on sphingolipids. Sphingosine-1-phosphate (S1P) is a lipid signaling mediator that functions both intracellularly, as a second messenger signaling molecule, and extracellularly, as a ligand for G-protein coupled S1P receptors (S1PRs). S1P plays important roles in modulating diverse cellular functions involved in cancer, i.e. cell growth, survival, and motility. Sphingosine Kinase 1 (SPHK1) catalyzes the phosphorylation of sphingosine, an amino alcohol, to generate S1P. Recently, studies have shown that SPHK1 is overexpressed in breast cancer and correlates with poor prognosis; thus, its role in breast cancer has received substantial attention. It is still unclear if SPHK1 plays any role in spontaneous metastasis of breast cancer to the lungs, and if so, the underlying mechanism behind it. To study this, we used the highly metastatic human triple negative breast cancer cell line MDA-MB-435, as it expressed high levels of endogenous SPHK1. We generated SPHK1 knockdown (435 shSPHK1) and control (435 shScramble) cells using SPHK1-targeting shRNA and scrambled shRNA, respectively. Knocking down SPHK1 decreased the invasive and migration potential of MDA-MB-435 cells in vitro, but had no significant effect on in vitro cell proliferation. Then, the control and knockdown cells were orthotopically injected into mammary fat pads (MFP) of nude mice to form MFP tumors. MFP tumors were excised from mice with survival surgery at day 30. Consistent with our in vitro proliferation data, there was no significant difference in MFP tumor proliferation, indicated by Ki-67 staining, between control and SPHK1 knockdown cells by day 30. However, TUNEL staining of MFP tumors showed that tumors formed by 435 shSPHK1 cells have increased apoptosis compared to tumors formed by 435 shScramble cells. After tumor excision at day 30, mice were monitored for another 4 weeks for spontaneous metastases. Remarkably, although ∼100% of mice injected with 435 shScramble developed spontaneous lung metastasis, only ∼20% of mice injected with 435 shSPHK1 had spontaneous lung metastasis. No metastasis was detected in the liver and the brain in this time frame. These data suggest that SPHK1 might play an important role in promoting spontaneous lung metastasis of breast cancer. Currently, we are investigating the specific step(s) of the metastatic cascade regulated by SPHK1. An understanding of the molecular mechanism of SPHK1-mediated breast cancer lung metastasis may enable SPHK1 to be developed as a molecular marker and a therapeutic target of metastatic breast cancer. Citation Format: Sunil Acharya, Chenyu Zhang, Frank L. Lowery, Qingling Zhang, Dihua Yu. SPHK1 promotes lung metastasis of breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2010. doi:10.1158/1538-7445.AM2014-2010

Inflammatory biomarker C-reactive protein and radiotherapy-induced early adverse skin reactions in patients with breast cancer.

Breast cancer is the most frequently diagnosed cancer and the second leading cause of cancer death in American women. Postsurgery adjuvant radiotherapy (RT) significantly reduced the local recurrence rate. However, many patients develop early adverse skin reactions (EASR) that impact quality of life and treatment outcomes. We evaluated an inflammatory biomarker, C-reactive protein (CRP), in predicting RT-induced EASRs in 159 patients with breast cancer undergoing RT. In each patient, we measured pre- and post-RT plasma CRP levels using a highly sensitive ELISA CRP assay. RT-induced EASRs were assessed at weeks 3 and 6 using the National Cancer Institute Common Toxicity Criteria (v3.0). Associations between EASRs and CRP levels were assessed using logistic regression models after adjusting for potential confounders. RT-induced grade 2+ EASRs were observed in 8 (5%) and 80 (50%) patients at weeks 3 and 6 (end of RT), respectively. At the end of RT, a significantly higher proportion of African Americans developed grade 3 EASRs (13.8% vs. 2.3% in others); grade 2+ EASRs were significantly associated with: change of CRP > 1 mg/L [odds ratio (OR), 2.51; 95% confidence interval (CI), 1.06-5.95; P = 0.04], obesity (OR, 2.08; 95% CI, 1.03-4.21; P = 0.04), or combined both factors (OR, 5.21; 95% CI, 1.77-15.38; P = 0.003). This is the first study to demonstrate that an inflammatory biomarker CRP is associated with RT-induced EASRs, particularly combined with obesity. Future larger studies are warranted to validate our findings and facilitate the discovery and development of anti-inflammatory agents to protect normal tissue from RT-induced adverse effects and improve quality of life in patients with breast cancer undergoing RT.

A Prospective Randomized Double-Blinded Trial of Topical NSAID Therapy for the Reduction of Pain in Women Receiving Breast Radiation for Breast Cancer

Breast cancer is the most common cancer and second leading cause of cancer death in American women. Whole breast radiation is given after mastectomy or lumpectomy to prevent breast cancer recurrence. Up to 95% of patients who undergo radiation therapy will experience an inflammatory-mediated skin reaction. Ketoprofen is a non-steroidal anti-inflammatory drug (NSAID) that helps decrease inflammation and pain by inhibiting prostaglandin synthesis. We hypothesize that topical ketoprofen will decrease discomfort in patients undergoing whole breast radiotherapy for breast cancer through this mechanism.